Two telomere-to-telomere gapless genomes reveal insights into Capsicum evolution and capsaicinoid biosynthesis.

Chili pepper (Capsicum) is known for its unique fruit pungency due to the presence of capsaicinoids. The evolutionary history of capsaicinoid biosynthesis and the mechanism of their tissue specificity remain obscure due to the lack of high-quality Capsicum genomes. Here, we report two telomere-to-telomere (T2T) gap-free genomes of C. annuum and its wild nonpungent relative C. rhomboideum to investigate the evolution of fruit pungency in chili peppers. We precisely delineate Capsicum centromeres, which lack high-copy tandem repeats but are extensively invaded by CRM retrotransposons. Through phylogenomic analyses, we estimate the evolutionary timing of capsaicinoid biosynthesis. We reveal disrupted coding and regulatory regions of key biosynthesis genes in nonpungent species. We also find conserved placenta-specific accessible chromatin regions, which likely allow for tissue-specific biosynthetic gene coregulation and capsaicinoid accumulation. These T2T genomic resources will accelerate chili pepper genetic improvement and help to understand Capsicum genome evolution.

FoPGDB: a pangenome database of Fusarium oxysporum, a cross-kingdom fungal pathogen.

Pangenomes, capturing the genetic diversity of a species or genus, are essential to understanding the ecology, pathobiology and evolutionary mechanisms of fungi that cause infection in crops and humans. However, fungal pangenome databases remain unavailable. Here, we report the first fungal pangenome database, specifically for Fusarium oxysporum species complex (FOSC), a group of cross-kingdom pathogens causing devastating vascular wilt to over 100 plant species and life-threatening fusariosis to immunocompromised humans. The F. oxysporum Pangenome Database (FoPGDB) is a comprehensive resource integrating 35 high-quality FOSC genomes, coupled with robust analytical tools. FoPGDB allows for both gene-based and graph-based exploration of the F. oxysporum pangenome. It also curates a large repository of putative effector sequences, crucial for understanding the mechanisms of FOSC pathogenicity. With an assortment of functionalities including gene search, genomic variant exploration and tools for functional enrichment, FoPGDB provides a platform for in-depth investigations of the genetic diversity and adaptability of F. oxysporum. The modular and user-friendly interface ensures efficient data access and interpretation. FoPGDB promises to be a valuable resource for F. oxysporum research, contributing to our understanding of this pathogen's pangenomic landscape and aiding in the development of novel disease management strategies. Database URL: http://www.fopgdb.site.

De novo chromosome-level genome assembly of Chinese motherwort (Leonurus japonicus).

Chinese motherwort (Leonurus japonicus), a member of Lamiaceae family, is a commonly used medicinal herb for treating obstetrical and gynecological diseases, producing over 280 officinal natural products. Due to limited genomic resources, little progress has been made in deciphering the biosynthetic pathway of valuable natural products in L. japonicus. Here, we de novo assembled the L. japonicus genome using high-coverage ONT long reads and Hi-C reads. The chromosome-level genome assembly contained ten chromosomes representing 99.29% of 489.34 Mb genomic sequence with a contig and scaffold N50 of 7.27 Mb and 50.86 Mb, respectively. Genome validations revealed BUSCO and LAI score of 99.2% and 21.99, respectively, suggesting high quality of genome assembly. Using transcriptomic data from various tissues, 22,531 protein-coding genes were annotated. Phylogenomic analysis of 13 angiosperm plants suggested L. japonicus had 58 expanded gene families functionally enriched in specialized metabolism such as diterpenoid biosynthesis. The genome assembly, annotation, and sequencing data provide resources for the elucidation of biosynthetic pathways behind natural products of pharmaceutical applications in L. japonicus.

Green ultrasonication-assisted extraction of microalgae Chlorella sp. for polysaturated fatty acid (PUFA) rich lipid extract using alternative solvent mixture.

Conventionally, microalgal lipid extraction uses volatile organic compounds as an extraction solvent. However, these solvents are harmful to human and environmental health. Therefore, this study evaluated the feasibility of alternative green solvents, namely, ethanol, dimethyl carbonate (DMC), cyclopentyl methyl ether (CPME), and 2-methyltetrahydrofuran (2-MeTHF) in lipid extraction from Chlorella sp. via ultrasound-assisted extraction (UAE). This study indicated that extraction parameters, such as ethanol-to-2-MeTHF ratio, solvent-to-biomass ratio, temperature, and time, significantly affected the crude lipid yield (P < 0.05). The highest crude lipid yield of 25.05 ± 0.924% was achieved using ethanol-2-MeTHF mixture (2:1, v/v) with a solvent-to-biomass ratio of 20:1 (v/w) at 60 °C for 25 min accompanying 100 W and 40 kHz. Ethanol-2-MeTHF-extracted lipids showed dominance in linoleic acid, α-linolenic acid, and palmitic acid. Overall this findings supported UAE using ethanol and 2-MeTHF as extraction solvents is a promising green alternative to conventional solvent extraction of lipids from microalgae.

Gapless genome assembly of Fusarium verticillioides, a filamentous fungus threatening plant and human health.

Fusarium verticillioides is a filamentous fungus that causes plant diseases and harms human health through cancer-inducing mycotoxin and life-threatening Fusariosis. Given its threat to agriculture and public health, genome assembly of this fungus is critical to our understanding of its pathobiology and developing antifungal drugs. Here, we report a gap-free genome assembly of F. verticillioides using PacBio HiFi data and high-throughput chromosome capture (Hi-C) sequencing data. The assembled 42.0 Mb sequence contains eleven gapless chromosomes capturing all centromeres and 19 of all 22 telomeres. This assembly represents a significant improvement over previous version on contiguity (contig N50: 4.3 Mb), completeness (BUSCO score: 99.0%) and correctness (QV: 88.8). A total of 15,230 protein-coding genes were predicted, 6.2% of which are newly annotated genes. In addition, we identified three-dimension chromatin structures such as TADs-like structures and chromatin loops based on Hi-C data of ultra-high coverage. This gap-free genome of F. verticillioides is an excellent resource for further panoramic understanding mechanisms of fungal genome evolution, mycotoxin production and pathogenesis on plant and human host.

Degradation of geosmin and 2-methylisoborneol in UV-based AOPs for photoreactors with reflective inner surfaces: Kinetics and transformation products.

Geosmin (GSM) and 2-methylisoborneol (2-MIB) are representative musty/earthy odor compounds commonly present in surface water. In present study, the degradation of GSM and 2-MIB subject to different UV-based advanced oxidation processes (AOPs), including UV/H2O2, UV/S2O82-, UV/chlorine, and UV/chloramine, in a phosphate-buffered saline (PBS) was conducted in a photoreactor with reflective inner surfaces and compared with that in an environmental water sample. A dynamic model to predict the degradation of GSM and 2-MIB in the photoreactor with reflective inner surfaces in the four UV-based AOPs was developed applying the second-order rate constants for the GSM and 2-MIB with primary reactive species (i.e., •OH, •Cl, and •SO4-) determined in this study. The model was proven to successfully simulate the degradation of GSM and 2-MIB. In addition, 8, 7, 8, and 11 degradation intermediates were detected from UV/H2O2, UV/S2O82-, UV/chlorine, and UV/chloramine in this study, and possible degradation pathways were proposed. This study is the first to report the degradation kinetics and formation products of GSM and 2-MIB in UV/chloramine. Research based on photoreactors with reflective inner surfaces may provide some guidance for eliminating GSM and 2-MIB in UV-based AOPs for full-scale engineering applications.

Analysis of microalgal growth kinetic model and carbohydrate biosynthesis cultivated using agro-industrial waste residuals as carbon source.

Microalgal carbohydrate is considered one of the potential feedstock for biofuel produced via the bioconversion process. However, the current cultivation process using commercial medium exhibited low biomass production and its carbohydrate productivity which become a bottleneck for sustainable microalgal-carbohydrate-based biofuel production. Thus, the objective of this study is to assess the utilization of industrial waste including molasses and glycerol on the Halochlorella rubescens and Tetraselmis suecica growth as well as its carbohydrate content under different cultivation modes such as autotrophic, heterotrophic and photoheterotrophic conditions. From this study, the highest maximum biomass of H. rubenscens and T. suecica of 0.653 ± 0.009 and 0.669 ± 0.01gL-1 were obtained when the cultivation was performed under photoheterotrophic using molasses. High carbohydrate content of H. rubescens and T. seucica of 56.81 ± 0.39% and 71.52 ± 0.03% with glucose represent the dominant sugar was observed under this condition. The growth kinetic model of the analysis indicated that Huang and Gompertz Models described well the growth of H. rubescens and T. suecica under photoheteroptroph condition with a high significant R2 of 0.99. The information generated could be beneficial for the future development of low-cost microalgal cultivation media formulation for future microalgal carbohydrate-based products such as bioethanol.

Atypical Ductal Hyperplasia of the Breast on Core Needle Biopsy: Risk of Malignant Upgrade on Surgical Excision / 한국유방암학회지

Purpose@#This study identified factors predicting malignant upgrade for atypical ductal hyperplasia (ADH) diagnosed on core-needle biopsy (CNB) and developed a nomogram to facilitate evidence-based decision making. @*Methods@#This retrospective analysis included women diagnosed with ADH at the National Cancer Centre Singapore (NCCS) in 2010–2015. Cox proportional hazards regression was used to identify clinical, radiological, and histological factors associated with malignant upgrade. A nomogram was constructed using variables with the strongest associations in multivariate analysis. Multivariable logistic regression coefficients were used to estimate the predicted probability of upgrade for each factor combination. @*Results@#Between 2010 and 2015, 238,122 women underwent mammographic screening under the National Breast Cancer Screening Program. Among 29,564 women recalled, 5,971 CNBs were performed. Of these, 2,876 underwent CNBs at NCCS, with 88 patients (90 lesions) diagnosed with ADH and 26 lesions upgraded to breast malignancy on excision biopsy. In univariate analysis, factors associated with malignant upgrade were the presence of a mass on ultrasound (p = 0.018) or mammography (p = 0.026), microcalcifications (p = 0.047), diffuse microcalcification distribution (p = 0.034), mammographic parenchymal density (p = 0.008). and ≥ 3 separate ADH foci found on biopsy (p = 0.024). Mammographic parenchymal density (hazard ratio [HR], 0.04; 95% confidence interval [CI], 0.005–0.35; p = 0.014), presence of a mass on ultrasound (HR, 10.50; 95% CI, 9.21–25.2; p = 0.010), and number of ADH foci (HR, 1.877; 95% CI, 1.831–1.920; p = 0.002) remained significant in multivariate analysis and were included in the nomogram. @*Conclusion@#Our model provided good discrimination of breast cancer risk prediction (C-statistic of 0.81; 95% CI, 0.74–0.88) and selected for a subset of women at low risk (2.1%) of malignant upgrade, who may avoid surgical excision following a CNB diagnosis of ADH.

Value of vascular endothelial-cadherin in evaluating the severity of patients with sepsis / 中华危重病急救医学

Objective:To explore the value of vascular endothelial-cadherin (VE-cad) in evaluating the severity of sepsis.Methods:A prospective study was conducted to select 85 patients with sepsis treated in the emergency ward of the First Affiliated Hospital of Wenzhou Medical University from June 1, 2015 to November 1, 2017. The gender, age, medical history, first infection site, number of affected organs, laboratory indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ), simplified acute physiology score Ⅱ(SAPSⅡ), sequential organ failure assessment (SOFA) and the total length of stay, emergency intensive care unit (EICU) length of stay, 28-day at admission and survival during hospitalization were measured, and the VE-cad level within 24 hours at admission was measured. The patients were divided into sepsis group and septic shock group according to the progress of the disease. The patients were divided into multiple organ dysfunction syndrome (MODS) group and non MODS group according to whether they were accompanied by MODS. The differences of the above indexes in patients with different disease progression, MODS and different prognosis were analyzed and compared. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the value of VE-cad in evaluating the severity of sepsis.Results:A total of 85 patients were included, mainly respiratory tract infection. Among them, 38 cases were sepsis and 47 cases were septic shock, 39 cases had MODS, 46 cases had no MODS, 64 cases survived and 21 cases died within 28 days after admission. Compared with sepsis group, the number of affected organs in septic shock group was greater [3 (2, 4) vs. 1 (0, 2)], APACHE Ⅱscore [13 (10, 21) vs. 7 (5, 12)], SAPS Ⅱscore [35 (31, 55) vs. 7 (5, 12)], SOFA score [7.0 (5.0, 10.0) vs. 3.0 (0, 5.0)], blood lactic acid [Lac (mmol/L): 3.5 (2.4, 6.2) vs. 1.9 (1.2, 2.2)], C-reactive protein [CRP (mg/L): 90.0 (58.1, 90.0) vs. 50.5 (38.0, 90.0)] and VE-cad levels [mg/L: 1.427 (1.141, 2.150) vs. 1.195 (0.901, 1.688)] were significantly increased, while platelet count [PLT (×10 9/L): 113.4±67.2 vs. 202.5±109.5] and hemoglobin (Hb) levels (g/L: 106.3±36.3 vs. 118.6±18.0) were significantly decreased (all P < 0.05). Compared with non MODS group, APACHE Ⅱ score [14 (10, 22) vs. 8 (6, 13)], SAPS Ⅱ score [36 (32, 56) vs. 29 (24, 35)], SOFA score (7.9±3.9 vs. 4.0±3.8), in-hospital mortality [53.8% (21/39) vs. 0% (0/46)], Lac [mmol/L: 3.1 (2.3, 6.3) vs. 2.1 (1.4, 4.6)] and VE-cad levels [mg/L: 1.427 (1.156, 1.937) vs. 1.195 (0.897, 1.776)] in MODS group were significantly higher, the length of stay in EICU was significantly longer [days: 6 (3, 12) vs. 3 (0, 7)], and the PLT level was significantly lower (×10 9/L: 118.2±80.0 vs. 182.5±104.0, all P < 0.05). Compared with the death group, the number of affected organs in the survival group was fewer [2 (1, 3) vs. 3 (1, 5)], APACHE Ⅱ score [9 (6, 13) vs. 21 (13, 25)], SAPS Ⅱ score [31 (25, 36) vs. 55 (35, 63)] and SOFA score (4.7±3.7 vs. 8.9±4.5) were significantly reduced, and the length of stay in EICU [days: 4 (1, 8) vs. 8 (3, 15)] was significantly shorter (all P < 0.05). ROC curve analysis showed that area under the ROC curve (AUC) of VE-cad, SOFA score and VE-cad combined with SOFA score in evaluating the severity of sepsis were 0.632 [95% confidence interval (95% CI) was 0.513-0.750], 0.830 (95% CI was 0.744-0.916) and 0.856 (95% CI was 0.779-0.933), respectively. When the cut-off value of VE-cad was 1.240 mg/L, the sensitivity was 68.1% and the specificity was 55.3%, the sensitivity of VE-cad combined with SOFA score was 85.1%, the specificity was 73.7%. Conclusion:VE-cad has a certain evaluation value for the severity of sepsis, and the evaluation value of VE-cad combined with SOFA score is better than that of VE-cad single index.

The optimal dose of oxidants in UV-based advanced oxidation processes with respect to primary radical concentrations.

UV-based advanced oxidation processes (AOPs) via photolysis of precursor chemical oxidants have been of interest to numerous researchers over the past several decades due to their capacity to generate highly active radical species and interesting radical chemistry. However, applications of UV-based AOPs have been commonly optimized case by case, due to the lack of theoretical investigations on process optimization, especially on oxidant doses. In this study, a simple equation for UV/H2O2 (•OH as the sole primary reactive species (PRS)) to obtain the theoretical optimal concentration (Copt-theoretical) for H2O2 was derived (Copt-theoretical=Ab·Scε·k). The equation was then validated for its accuracy in the calculation of Copt-theoretical for H2O2 in the UV/H2O2 AOP using a well-established comprehensive kinetic model. A competition kinetics method for the measurement of scavenging capacity (Sc, the unknown parameter for the simple equation) was designed, for which nitrobenzene was employed as the probe compound and tert­butyl alcohol was introduced as the standard compound. Based on this simple equation, we calculated the Copt-theoretical of 77 environmental water samples and introduced the concept of a practical optimal oxidants dose for the UV/H2O2 AOP, while minimizing the operation costs in engineering applications. Moreover, this study mathematically proved that the simple equation obtained from UV/H2O2 could be successfully extended to other UV-based AOPs, including UV/chlorine, UV/NH2Cl, UV/S2O82-, and UV/peracetic acid. The simple equation of Copt-theoretical derived in this study may not only help to provide instructions for engineering applications, but also point out the ultimate treatment capability of each UV-based AOPs.

Maternal bone adaptation to mechanical loading during pregnancy, lactation, and post-weaning recovery.

The maternal skeleton undergoes dramatic bone loss during pregnancy and lactation, and substantial bone recovery post-weaning. The structural adaptations of maternal bone during reproduction and lactation exert a better protection of the mechanical integrity at the critical load-bearing sites, suggesting the importance of physiological load-bearing in regulating reproduction-induced skeletal alterations. Although it is suggested that physical exercise during pregnancy and breastfeeding improves women's physical and psychological well-being, its effects on maternal bone health remain unclear. Therefore, the objective of this study was to investigate the maternal bone adaptations to external mechanical loading during pregnancy, lactation, and post-weaning recovery. By utilizing an in vivo dynamic tibial loading protocol in a rat model, we demonstrated improved maternal cortical bone structure in response to dynamic loading at tibial midshaft, regardless of reproductive status. Notably, despite the minimal loading responses detected in the trabecular bone in virgins, rat bone during lactation experienced enhanced mechano-responsiveness in both trabecular and cortical bone compartments when compared to rats at other reproductive stages or age-matched virgins. Furthermore, our study showed that the lactation-induced elevation in osteocyte peri-lacunar/canalicular remodeling (PLR) activities led to enlarged osteocyte lacunae. This may result in alterations in interstitial fluid flow-mediated mechanical stimulation on osteocytes and an elevation in solute transport through the lacunar-canalicular system (LCS) during high-frequency dynamic loading, thus enhancing mechano-responsiveness of maternal bone during lactation. Taken together, findings from this study provide important insights into the relationship between reproduction- and lactation-induced skeletal changes and external mechanical loading, emphasizing the importance of weight-bearing exercise on maternal bone health during reproduction and postpartum.

Bacillus megaterium: a Potential and an Efficient Bio-Degrader of Polystyrene

Abstract This research has been conducted to investigate the biodegradation of polystyrene (PS) by isolated strain Bacillus megaterium from Zophobas morio's. The viability and metabolic activity of Bacillus megaterium utilizing emulsified PS in liquid carbon free basal medium as sole carbon source was confirmed using redox probe 2,3,5-triphenyltetrazolium chloride (TTC). Bacillus megaterium showed prominent result with TTC, which forms red colored insoluble TPF (triphenyl formazan) within four to seven days. The formation of TPF confirmed the metabolism activity of the cell using PS as source of carbon. The biodegradation activity of Bacillus megaterium on PS film was also confirmed by FTIR analysis, the result showed changes in functional groups due to microbial activities. FESEM analysis was carried out on the PS surface, revealing the formation of bacterial biofilm as well as occurrence of porosity and fragility of the bacterial deteriorate surface compared to control. Overall, the identification of plastic degrading bacteria (PDB) will become a fundamental platform promoting more in-depth research for optimal plastic waste management to switch from the conventional landfilling to enzymatic biodegradation. In the long run, it is to regulate the current global plastic waste accumulation on Earth which poses potent ecological threat.

Achieving GWAS with homomorphic encryption.

BACKGROUND: One way of investigating how genes affect human traits would be with a genome-wide association study (GWAS). Genetic markers, known as single-nucleotide polymorphism (SNP), are used in GWAS. This raises privacy and security concerns as these genetic markers can be used to identify individuals uniquely. This problem is further exacerbated by a large number of SNPs needed, which produce reliable results at a higher risk of compromising the privacy of participants. METHODS: We describe a method using homomorphic encryption (HE) to perform GWAS in a secure and private setting. This work is based on a proposed algorithm. Our solution mainly involves homomorphically encrypted matrix operations and suitable approximations that adapts the semi-parallel GWAS algorithm for HE. We leverage upon the complex space of the CKKS encryption scheme to increase the number of SNPs that can be packed within a ciphertext. We have also developed a cache module that manages ciphertexts, reducing the memory footprint. RESULTS: We have implemented our solution over two HE open source libraries, HEAAN and SEAL. Our best implementation took 24.70 minutes for a dataset with 245 samples, over 4 covariates and 10643 SNPs. CONCLUSIONS: We demonstrate that it is possible to achieve GWAS with homomorphic encryption with suitable approximations.

Reactive Nitrogen Species Mediated Degradation of Estrogenic Disrupting Chemicals by Biochar/Monochloramine in Buffered Water and Synthetic Hydrolyzed Urine.

There is increasing concern about the severe endocrine-related health problems because of the discharge of estrogenic disrupting chemicals (EDCs) into the natural environment. In this study, we investigated the activation of monochloramine (NH2Cl) by biochar [pyrolyzed by cotton straw at 350 °C (Cot350), wheat straw at 350 and 700 °C (WS350 and WS700), and corn straw at 350 and 700 °C (CS350 and CS700)] for the degradation of estradiol (E2) and ethinylestradiol (EE2). Approximately 95% of parent E2 and EE2 was removed by Cot350/NH2Cl in buffered solution, and 87% of E2 and 75% of EE2 were removed in urine within 24 h. Electronic paramagnetic resonance analysis and radical-quenching experiments showed that biochar activated NH2Cl and primarily generated •NO radicals for the degradation of the EDCs. The nitrogen and silicon elements of Cot350 served as primary catalytic sites for NH2Cl activation, whereas the sp2-hybridized carbon on WS700 and CS700 played a major role. The effect of major urine components (i.e., ammonia species, chloride, and bicarbonate) on the reaction pathways of biochar/NH2Cl was also elucidated. This study provides new insights into the reaction pathways of NH2Cl activation by biochar and suggests potential applications for other carbonaceous materials for NH2Cl activation.

Degradation of Organic Micropollutants in UV/NH2Cl Advanced Oxidation Process.

Monochloramine (NH2Cl) can be irradiated by UV to create an advanced oxidation condition (i.e., UV/NH2Cl) for the elimination of organic micropollutants (OMPs) from source water. However, information in retrospective studies was scarce on how UV/NH2Cl performance would be affected by the water matrix and OMP molecular structures. In this study, the degradation of five representative OMPs, including triclosan, carbamazepine, sulfamethoxazole, estradiol (E2), and ethinylestradiol (EE2), was examined in different water matrices. All OMPs were rapidly removed by UV/NH2Cl but exhibited different degradation mechanisms. Although •OH, •Cl, and direct photolysis mainly contributed to the overall degradation of OMPs in buffered nanopure water, the contribution of reactive nitrogen species (RNS) generated from the photolysis of NH2Cl was not negligible in the degradation of E2 and EE2. A phenolic group was identified as the moiety reactive toward RNS. Based on quantitative analysis of the impact on OMP degradation from cosolutes (including Cl-, HCO3-, NOM) as well as pH and NH2Cl doses, we developed a kinetic model for the prediction of OMP degradation in complex water matrices. In environmental water matrices, the performance and radical contributions in UV/NH2Cl and UV/H2O2 systems were taken into comparison, which showed faster degradation of OMPs and a more significant contribution of CO3•- in the UV/NH2Cl process.

A web-based educational intervention module to improve knowledge and attitudes towards thalassaemia prevention in Malaysian young adults

@#Background: Thalassaemia is a public health burden in Malaysia and its prevention faces many challenges. In this study, we aimed to assess the effectiveness of a web-based educational module in improving knowledge and attitudes about thalassaemia prevention amongst Malaysian young adults. Methods: We designed an interactive web-based educational module in the Malay language wherein videos were combined with text and pictorial visual cues. Malaysians aged 18-40 years old who underwent the module had their knowledge and attitudes assessed at baseline, post-intervention and at 6-month follow-up using a selfadministered validated questionnaire. Results: Sixty-five participants: 47 Malays (72.3%), 15 Chinese (23.1%), three Indians (4.6%) underwent the module. Questionnaires were completed at baseline (n=65), postintervention (n=65) and at 6-month follow-up (n=60). Out of a total knowledge score of 21, significant changes were recorded across three time-points- median scores were 12 at pre-intervention, 19 at post-intervention and 16 at 6-month follow-up (p<0.001). Post-hoc testing comparing preintervention and 6-month follow-up scores showed significant retention of knowledge (p<0.001). Compared to baseline, attitudes at 6-month follow-up showed an increased acceptance for “marriage avoidance between carriers” (pre-intervention 20%, 6-month follow-up 48.3%, p<0.001) and “prenatal diagnosis” (pre-intervention 73.8%, 6-month follow-up 86.2%, p=0.008). Acceptance for selective termination however, remained low without significant change (pre-intervention 6.2%, 6-month follow-up 16.7%, p=0.109). Conclusion: A web-based educational module appears effective in improving knowledge and attitudes towards thalassaemia prevention and its incorporation in thalassaemia prevention programs is potentially useful in Malaysia and countries with a high internet penetration rate.

Establishment of rat model of anti-glomerular basement membrane nephritis induced by pCol (24-38) / 解放军医学杂志

Objective Polypeptide pCol (24-38) containing rat non-collagen region of collagen type IV alpha 3 chain [α3(IV)NC1] was used as antigen to establish the rat model of anti-glomerular basement membrane (GBM) disease, and study the possible pathogenesis of anti-GBM disease. Methods Twenty male WKY rats, 8-10 weeks old and weight 120-150g, were randomly divided into control group and anti-GBM model group (10 each). Polypeptide pCol(24-38) was used as antigen in anti-GBM model group, while pCol (38-52) was used as antigen in control group. After emulsification with complete Freund's adjuvant, rats in the both groups were immunized by subcutaneous injection at three points on the back. 24-hour urine was collected regularly every week after immunization to measure the urinary protein content. All animals were sacrificed 49 days after immunization, and serum was taken. The serum creatinine level was detected by creatine oxidase method, urea nitrogen concentration was determined by urease method, and the relative titer of serum anti-pCol (24-38) antibodies was detected by indirect ELISA. Part of kidney tissue was stained with PAS and immunohistochemically treatment in paraffin section, and the other part was frozen section after OCT embedding to detect IgG deposition in glomeruli by direct immunofluorescence method. Results The 24-hour urinary protein levels increased significantly from the 4th week in anti-GBM model group than in control group [(52.27±10.50)mg/24h vs. (4.87±0.64)mg/24h, P<0.001], and then rose gradually until the 7th week [(255.80±9.79)mg/24h vs. (5.78±0.39)mg/24h, P<0.001]. Both the serum creatinine level and urea nitrogen level increased obviously in anti-GBM model group than in control group [(145.3±22.60)μmol/L vs. (36.81±2.21)μmol/L; (26.59±5.01)mmol/L vs. (6.92±0.27)mmol/L] with statistically significant difference (P<0.001). Compared with the control group, rats in anti-GBM model group produced high levels of circulating anti-pCol (24-38) IgG antibodies [(1.59±0.18) vs. (0.09±0.01)] with statistically significant difference (P<0.05). PAS staining showed glomerular sclerosis and diffuse crescent formation in anti-GBM model group; immunohistochemistry showed macrophage infiltration in glomerular; fluorescence staining showed obvious linear deposition of IgG in glomerular basement membrane. However, in the control group, PAS staining showed no abnormal glomerular, immunohistochemistry showed no macrophage infiltration, and fluorescence staining showed no IgG deposits on the glomerular basement membrane. Conclusions The animal model of anti-GBM disease may be successfully constructed by using polypeptide pCol (24-38) as antigen to immunize WKY rats. The preparation method of the antigen is simple, and so is valuable for designing new therapeutic strategies against anti-GBM disease.

Generalized polyspike train: An EEG biomarker of drug-resistant idiopathic generalized epilepsy.

OBJECTIVE: To identify clinical and EEG biomarkers of drug resistance in adults with idiopathic generalized epilepsy. METHODS: We conducted a case-control study consisting of a discovery cohort and a replication cohort independently assessed at 2 different centers. In each center, patients with the idiopathic generalized epilepsy phenotype and generalized spike-wave discharges on EEG were classified as drug-resistant or drug-responsive. EEG changes were classified into predefined patterns and compared between the 2 groups in the discovery cohort. Factors associated with drug resistance in multivariable analysis were tested in the replication cohort. RESULTS: The discovery cohort included 85 patients (29% drug-resistant and 71% drug-responsive). Their median age at assessment was 32 years and 50.6% were female. Multivariable analysis showed that higher number of seizure types ever experienced (3 vs 1: odds ratio [OR] = 31.1, 95% confidence interval [CI]: 4.5-214, p < 0.001; 3 vs 2: OR = 14.6, 95% CI: 2.3-93.1, p = 0.004) and generalized polyspike train (burst of generalized rhythmic spikes lasting less than 1 second) during sleep were associated with drug resistance (OR = 10.8, 95% CI: 2.4-49.4, p = 0.002). When these factors were tested in the replication cohort of 80 patients (27.5% drug-resistant and 72.5% drug-responsive; 71.3% female; median age 27.5 years), the proportion of patients with generalized polyspike train during sleep was also higher in the drug-resistant group (OR = 4.0, 95% CI: 1.35-11.8, p = 0.012). CONCLUSION: Generalized polyspike train during sleep may be an EEG biomarker for drug resistance in adults with idiopathic generalized epilepsy.